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. 2018 Apr 3;9(25):17512–17522. doi: 10.18632/oncotarget.24776

Figure 2. Effect of HNF-1β or Claspin on the expression of p-Chk1 and Claspin protein.

Figure 2

HNF-1β-knocked down cells were treated with bleomycin for 24 hours and p-Chk1 and Claspin were then quantified. High knock-down efficiency (85%) by si-HNF-1β was observed 24 h after bleomycin treatment (data not shown). (A and B) Effect of HNF-1β on the expression of p-Chk1 and Claspin protein in response to a genotoxic agent bleomycin. Protein levels of p-Chk1 and Claspin were compared among two (si-HNF-1β and si-Ctl) groups, with β-actin as an internal control. HNF-1β does not affect the expression of total Chk1 [7]. HNF-1β promoted p-Chk1 and Claspin expression for at least 24 hours. The protein levels of p-Chk1 and Claspin are coordinately overexpressed in response to HNF-1β. Expression of p-Chk1 and Claspin proteins was higher in TOV21G si-Ctl cells for 9–24 hours after bleomycin compared with TOV21G si-HNF-1β cells. (C) Effect of Claspin on the p-Chk1 protein expression after bleomycin. This figure shows the time-dependent expression of p-Chk1 protein in TOV21G si-Claspin cells or si-Ctl cells after bleomycin treatment. Figure shows representative images from at least three independent Western blot assays.